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This book presents the advances in super-resolution microscopy in physics and biomedical optics for nanoscale imaging. In the last decade, super-resolved fluorescence imaging has opened new horizons in improving the resolution of optical microscopes far beyond the classical diffraction limit, leading to the Nobel Prize in Chemistry in 2014. This book represents the first comprehensive review of a different type of super-resolved microscopy, which does not rely on using fluorescent markers. Such label-free super-resolution microscopy enables potentially even broader applications in life sciences and nanoscale imaging, but is much more challenging and it is based on different physical concepts and approaches. A unique feature of this book is that it combines insights into mechanisms of label-free super-resolution with a vast range of applications from fast imaging of living cells to inorganic nanostructures. This book can be used by researchers in biological and medical physics. Due to its logically organizational structure, it can be also used as a teaching tool in graduate and upper-division undergraduate-level courses devoted to super-resolved microscopy, nanoscale imaging, microscopy instrumentation, and biomedical imaging.
This book encompasses the full breadth of the super-resolution imaging field, representing modern techniques that exceed the traditional diffraction limit, thereby opening up new applications in biomedicine. It shows readers how to use the new tools to increase resolution in sub-nanometer-scale images of living cells and tissue, which leads to new information about molecules, pathways and dynamics. The book highlights the advantages and disadvantages of the techniques, and gives state-of-the-art examples of applications using microscopes currently available on the market. It covers key techniques such as stimulated emission depletion (STED), structured illumination microscopy (SSIM), photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM). It will be a useful reference for biomedical researchers who want to work with super-resolution imaging, learn the proper technique for their application, and simultaneously obtain a solid footing in other techniques.
This book describes developments in the field of super-resolution fluorescence microscopy or nanoscopy. In 11 chapters, distinguished scientists and leaders in their respective fields describe different nanoscopy approaches, various labeling technologies, and concrete applications. The topics covered include the principles and applications of the most popular nanoscopy techniques STED and (f)PALM/STORM, along with advances brought about by fluorescent proteins and organic dyes optimized for fluorescence nanoscopy. Furthermore, the photophysics of fluorescent labels is addressed, specifically for improving their photoswitching capabilities. Important applications are also discussed, such as the tracking and counting of molecules to determine acting forces in cells, and quantitative cellular imaging, respectively, as well as the mapping of chemical reaction centers at the nano-scale. The 2014 Chemistry Nobel Prize® was awarded for the ground-breaking developments of super-resolved fluorescence microscopy. In this book, which was co-edited by one of the prize winners, readers will find the most recent developments in this field.
The combination of electron microscopy with transmitted light microscopy (termed correlative light and electron microscopy; CLEM) has been employed for decades to generate molecular identification that can be visualized by a dark, electron-dense precipitate. This new volume of Methods in Cell Biology covers many areas of CLEM, including a brief history and overview on CLEM methods, imaging of intermediate stages of meiotic spindle assembly in C. elegans embryos using CLEM, and capturing endocytic segregation events with HPF-CLEM. Covers many areas of CLEM by the best international scientists in the field Includes a brief history and overview on CLEM methods
Stimulated Raman Scattering Microscopy: Techniques and Applications describes innovations in instrumentation, data science, chemical probe development, and various applications enabled by a state-of-the-art stimulated Raman scattering (SRS) microscope. Beginning by introducing the history of SRS, this book is composed of seven parts in depth including instrumentation strategies that have pushed the physical limits of SRS microscopy, vibrational probes (which increased the SRS imaging functionality), data science methods, and recent efforts in miniaturization. This rapidly growing field needs a comprehensive resource that brings together the current knowledge on the topic, and this book does just that. Researchers who need to know the requirements for all aspects of the instrumentation as well as the requirements of different imaging applications (such as different types of biological tissue) will benefit enormously from the examples of successful demonstrations of SRS imaging in the book. Led by Editor-in-Chief Ji-Xin Cheng, a pioneer in coherent Raman scattering microscopy, the editorial team has brought together various experts on each aspect of SRS imaging from around the world to provide an authoritative guide to this increasingly important imaging technique. This book is a comprehensive reference for researchers, faculty, postdoctoral researchers, and engineers. - Includes every aspect from theoretic reviews of SRS spectroscopy to innovations in instrumentation and current applications of SRS microscopy - Provides copious visual elements that illustrate key information, such as SRS images of various biological samples and instrument diagrams and schematics - Edited by leading experts of SRS microscopy, with each chapter written by experts in their given topics
This unique book on super-resolution microscopy techniques presents comparative, in-depth analyses of the strengths and weaknesses of the individual approaches. It was written for non-experts who need to understand the principles of super-resolution or who wish to use recently commercialized instruments as well as for professionals who plan to realize novel microscopic devices. Explaining the practical requirements in terms of hardware, software and sample preparation, the book offers a wealth of hands-on tips and practical tricks to get a setup running, provides invaluable help and support for successful data acquisition and specific advice in the context of data analysis and visualization. Furthermore, it addresses a wide array of transdisciplinary fields of applications. The author begins by outlining the joint efforts that have led to achieving super-resolution microscopy combining advances in single-molecule photo-physics, fluorophore design and fluorescent labeling, instrument design and software development. The following chapters depict and compare current main standard techniques such as structured illumination microscopy, single-molecule localization, stimulated emission depletion microscopy and multi-scale imaging including light-sheet and expansion microscopy. For each individual approach the experimental setups are introduced, the imaging protocols are provided and the various applications illustrated. The book concludes with a discussion of future challenges addressing issues of routine applications and further commercialization of the available methods. Guiding users in how to make choices for the design of their own experiments from scratch to promising application, this one-stop resource is intended for researchers in the applied sciences, from chemistry to biology and medicine to physics and engineering.
This book demonstrates the concept of Fourier ptychography, a new imaging technique that bypasses the resolution limit of the employed optics. In particular, it transforms the general challenge of high-throughput, high-resolution imaging from one that is coupled to the physical limitations of the optics to one that is solvable through computation. Demonstrated in a tutorial form and providing many MATLAB® simulation examples for the reader, it also discusses the experimental implementation and recent developments of Fourier ptychography. This book will be of interest to researchers and engineers learning simulation techniques for Fourier optics and the Fourier ptychography concept.
This book provides in depths information on different microscopy approaches and supplies the reader with methods how to untangle highly complex processes involved in physiological and pathophysiological cardiac signaling. Microscopy approaches have established themselves as the quasi gold standard that enables us to appreciate the underlying mechanisms of physiological and pathophysiological cardiac signaling. This book presents the most important microscopy techniques from the level of individual molecule e.g. Förster-Resonance Energy Transfer (FRET), up to cellular and tissue imaging, e.g. electron microscopy (TEM) or light sheet microscopy. The book is intended for graduate students and postdocs in cardiovascular research, imaging and cell biology, pre-clinical and clinical researchers in cardiovascular sciences as well as decision makers of the pharmaceutical industry.
The topics range from single molecule experiments in quantum optics and solid-state physics to analogous investigations in physical chemistry and biophysics.
This open access book, edited and authored by a team of world-leading researchers, provides a broad overview of advanced photonic methods for nanoscale visualization, as well as describing a range of fascinating in-depth studies. Introductory chapters cover the most relevant physics and basic methods that young researchers need to master in order to work effectively in the field of nanoscale photonic imaging, from physical first principles, to instrumentation, to mathematical foundations of imaging and data analysis. Subsequent chapters demonstrate how these cutting edge methods are applied to a variety of systems, including complex fluids and biomolecular systems, for visualizing their structure and dynamics, in space and on timescales extending over many orders of magnitude down to the femtosecond range. Progress in nanoscale photonic imaging in Göttingen has been the sum total of more than a decade of work by a wide range of scientists and mathematicians across disciplines, working together in a vibrant collaboration of a kind rarely matched. This volume presents the highlights of their research achievements and serves as a record of the unique and remarkable constellation of contributors, as well as looking ahead at the future prospects in this field. It will serve not only as a useful reference for experienced researchers but also as a valuable point of entry for newcomers.